1. Field of the Invention
The invention relates to glycoprotein micelles and multilayer glycoprotein vesicles. The invention further relates to methods of using glycoprotein micelles and glycoprotein vesicles to detect, concentrate and/or capture other molecular entities, such as biological contaminants, including pathogens and/or biotoxins, in the environment and to detect protein glycosylation.
2. Background Art
A major concern for municipal and commercial water treatment facilities is the detection of pathogenic microorganisms. For example, a number of chlorine resistant pathogens such as Crypotosporidium can contaminate drinking water systems and other potentially harmful microorganisms and/or biotoxins can be introduced either accidentally or intentionally.
For example, water supplies and water distributions systems represent potential targets for terrorist activity in the United States and other parts of the world because of the critical need for water in every sector of an industrialized society. Meinhardt, Ann. Rev. Pub. Health 26:213-237 (2005). Even short-term disruption of water service can significantly impact a community because of accidental or intentional contamination of a municipal water system as part of, for example a terrorist attack. Such contamination could lead to serious medical, public health, and economic consequences. Id.
In the past, people in the United States have largely taken for granted the convenience of potable municipal water. However, the threat of intentional contamination of our water supplies is becoming a concern because of a rise in the number of terrorist acts around the world. As a result, there is much interest in technologies that can be used to detect a contamination event in real time, or close to real time, as well as dispel or confirm the credibility of a threat. Such technologies include immunoassay devices that can be used to determine the presence of biotoxins and pathogens in water. Immunoassay devices are based on immunological interactions during which specific antibodies react with contaminants, or antigens, to produce a response indicating the presence of the contaminant. Examples of such devices include, BADDT Test Strips (ADVNT Biotechnologies), BioVerify Test Kits (BioVeris), EzyBot® A and EzyBot® B Test Kits (Pharmaleads), RAMP® Test Cartridges (Response Biomedical Corp.), BioThreat Alert® Test Strips (Tetracore, Inc.), Enzyme Linked Immunosorbent Assay (Tetracore, Inc.), and QTL Biosensor (QTL Biosystems LLC).
Such immunoassay devices can be expensive because each requires the presence of antibodies. Antibodies can be time consuming and difficult to make, requiring numerous process and purification steps and typically require the use of live animals. Moreover, the U.S. Environmental Protection Agency (“EPA”) website indicates that the EPA has tested a variety of the immunoassay devices and reports that several of the devices yielded false positive and false negative responses when testing for anthrax, botulinum toxin A and ricin. Additional biological contaminant detection techniques include the use of polymerase chain reaction, which is also costly and time consuming.
As noted, field-deployable detection technologies in the nation's water supplies have become a high priority in recent years. Biological monitoring devices can assess the type and extent of contamination in a suspected water security event. According to a May 17, 2007 ScienceDaily article, Sandia National Laboratories has developed a unattended water sensor (“UWS”) (measuring 17 inches high by 14 inches wide by 7 inches deep) in a box composed of analytic instruments, pumps, tubes, and small reservoirs to handle minute amounts of fluid. The reservoirs contain chemical buffers, fluorescent dyes, proteins, and separation gel. The diagnostic instrumentation package, based on Sandia's MicroChemLab technology, is mounted near the water supply. The box is connected to a small, submerged probe that transports the sample into the system.
Although the UWS is currently able to detect protein toxins such as SEB, botulinum, and ricin, the device cannot detect pathogens, such as bacteria, e.g., E. coli and protozoa such as Cryptosporidium. A working database of organism signatures must be developed to allow the device to accurately distinguish the signatures from one another.
The background of U.S. Patent Publication Number 2007/0195324 provides a historical perspective on systems and methods for detecting radiation, biotoxin, chemical and biological warfare agents. The publication also describes an optical particle detection system to identify and classify particles by capturing digitized images of the particle generated by directing a light source through a fluid that includes the particle. The particle scatters the light and the scattered light is detected. The bio-optical signature of the particle is then used to classify the event or particle. A count rate of the classified particles is monitored to detect a change that is representative of a toxin attack.
Despite the presence of techniques and devices for the detection of contaminants in water, there is still a need for inexpensive and simple field techniques for the detection of a variety of biological contaminants in water.